College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, China.
Jiangsu Simovay Pharmaceutical Co., Ltd, Nanjing 210042, China.
Phytomedicine. 2021 Mar;83:153469. doi: 10.1016/j.phymed.2021.153469. Epub 2021 Jan 17.
Ischemic stroke is a multifactorial disease contributing to mortality and neurological dysfunction. Isoliquiritin (ISL) has been reported to possess a series of pharmacological activities including antioxidant, anti-inflammatory, antifungal, anti-depression, anti-neurotoxicity and pro-angiogenesis activities but whether it can be used for ischemic stroke treatment remains unknown.
The goal of this study is to explore its therapeutic effect on ischemic stroke and demonstrated the potential mechanism of ISL in zebrafish model.
Using the photothrombotic-induced adult zebrafish model of ischemic stroke, we visualized the telencephalon (Tel) and optic tectum (OT) infarction injury at 24 h post-light exposure for 30 min by TTC and H&E staining. The effect of ISL on neurological deficits was analyzed during open tank swimming by video tracking. The antioxidant activity against ischemia injury was quantified by SOD, GSH-Px and MDA assay. Transcriptome analysis of zebrafish Tel revealed how ISL regulating gene expression to exert protective effect, which were also been validated by real-time quantitative PCR assays.
We found for the first time that the Tel tissue was the first damaged site of the whole brain and it showed more sensitivity to the brain ischemic damage compared to the OT. ISL reduced the rate of Tel injury, ameliorated neurological deficits as well as counteracted oxidative damages by increasing SOD, GSH-Px and decreasing MDA activity. GO enrichment demonstrated that ISL protected membrane and membrane function as well as initiate immune regulation in the stress response after ischemia. KEGG pathway analysis pointed out that immune-related pathways, apoptosis as well as necroptosis pathways were more involved in the protective mechanism of ISL. Furthermore, the log2 fold change in expression pattern of 25 genes detected by qRT-PCR was consistent with that by RNA-seq.
Tel was highly sensitive to the brain ischemia injury in zebrafish model of ischemic stroke. ISL significantly exerted protective effect on Tel injury, neurological deficits and oxidative damages. ISL could regulate a variety of genes related to immune, apoptosis and necrosis pathways against complex cascade reaction after ischemia. These findings enriched the study of ISL, making it a novel multi-target agent for ischemic stroke treatment.
缺血性脑卒中是一种多因素疾病，可导致死亡率和神经功能障碍。已报道甘草查尔酮 B（ISL）具有一系列药理活性，包括抗氧化、抗炎、抗真菌、抗抑郁、抗神经毒性和促血管生成作用，但它是否可用于治疗缺血性脑卒中尚不清楚。
本研究旨在探讨其在缺血性脑卒中治疗中的疗效，并在斑马鱼模型中阐明 ISL 的潜在作用机制。
利用光血栓诱导的成年斑马鱼缺血性脑卒中模型，通过 TTC 和 H&E 染色在光暴露 30 分钟后 24 小时可视化端脑（Tel）和视顶盖（OT）梗死损伤。通过视频跟踪分析 ISL 对开放泳槽中神经功能缺损的影响。通过 SOD、GSH-Px 和 MDA 测定来量化 ISL 对缺血损伤的抗氧化活性。通过实时定量 PCR 检测对斑马鱼 Tel 的转录组分析揭示了 ISL 如何调节基因表达以发挥保护作用。
我们首次发现 Tel 组织是全脑的第一个受损部位，与 OT 相比，它对脑缺血损伤更为敏感。ISL 降低了 Tel 损伤率，改善了神经功能缺损，并通过增加 SOD、GSH-Px 和降低 MDA 活性来对抗氧化损伤。GO 富集表明，ISL 在缺血后应激反应中保护了膜和膜功能，并启动了免疫调节。KEGG 通路分析指出，免疫相关通路、细胞凋亡和坏死通路更多地参与了 ISL 的保护机制。此外，qRT-PCR 检测到的 25 个基因的表达模式的 log2 倍变化与 RNA-seq 一致。
Tel 在斑马鱼缺血性脑卒中模型中对脑缺血损伤高度敏感。ISL 对 Tel 损伤、神经功能缺损和氧化损伤具有显著的保护作用。ISL 可调节与免疫、细胞凋亡和坏死通路相关的多种基因，以应对缺血后的复杂级联反应。这些发现丰富了 ISL 的研究，使其成为一种新型的缺血性脑卒中治疗多靶点药物。
